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Figure 3


Fig. 3. Cdc42–Par-6 interaction is necessary for neuroblast polarity. (A) Alignment of the Par-6 semi-CRIB domain with CRIB domains from other proteins. Mutated residues are boxed and the residues mutated in the Par-6ISAA transgene are boxed in red. (B) The ISAA mutation disrupts Cdc42 binding to the Par-6 CRIB-PDZ domain. The extent of binding between a glutathione-S-transferase (GST) fusion of [{gamma}35S]GTP-loaded Cdc42 and 55 µM wild-type and mutant Par-6 CRIB-PDZ domains is shown, as determined using a qualitative pull-down assay stained with Coomassie brilliant blue. (C,D) Zygotic par6{Delta}226 central brain neuroblasts 24 hours ALH expressing par-6 transgenes. HA–Par-6 (HA:Par-6) localizes to the apical cortex of dividing neuroblasts and rescues Mira phenotype (C). HA–Par-6ISAA (HA:Par-6ISAA) is cytoplasmic and is unable to rescue cortical Mira (D). (E) Zygotic par6{Delta}226 central brain neuroblasts 24 hours ALH expressing Cdc42-Myc. Arrowhead delineates weak apical enrichment of Cdc42-Myc (92%, n=12), whereas Mira is uniformally cortical (100%, n=12).





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